Photoinduced Dynamics of 13,13'-Diphenylpropyl-ß-carotene.

Carotenoids are ubiquitous pigment systems in nature which are relevant to a range of processes, such as photosynthesis, but the detailed influence of substitutions at the polyene backbone on their photophysics is still underexplored. Here, we present a detailed experimental and theoretical investigation of the carotenoid 13,13'-diphenylpropyl-ß-carotene using ultrafast transient absorption spectroscopy and steady-state absorption experiments in n-hexane and n-hexadecane, complemented by DFT/TDDFT calculations. In spite of their bulkiness and their potential capability to "fold back" onto the polyene system, which could result in π-stacking effects, the phenylpropyl residues have only a minor impact on the photophysical properties compared with the parent compound ß-carotene. Ultrafast spectroscopy finds lifetimes of 200-300 fs for the S2 state and 8.3-9.5 ps for the S1 state. Intramolecular vibrational redistribution with time constants in the range 0.6-1.4 ps is observed in terms of a spectral narrowing of the S1 spectrum over time. We also find clear indications of the presence of vibrationally hot molecules in the ground electronic state (S0*). The DFT/TDDFT calculations confirm that the propyl spacer electronically decouples the phenyl and polyene π-systems and that the substituents in the 13 and 13' positions point away from the polyene system.

Determination of Hydrophobic Dispersive Surface Free Energy of Activated Carbon Fibers Measured by Inverse Gas Chromatographic Technique.

Activated carbon fibers (ACFs) as one of the most important porous carbon materials are widely used in many applications that involve rapid adsorption and low-pressure loss, including air purification, water treatment, and electrochemical applications. For designing such fibers for the adsorption bed in gas and aqueous phases, in-depth comprehension of the surface components is crucial. However, achieving reliable values remains a major challenge due to the high adsorption affinity of ACFs. To overcome this problem, we propose a novel approach to determine London dispersive components (γSL) of the surface free energy of ACFs by inverse gas chromatography (IGC) technique at an infinite dilution. Our data reveal the γSL values at 298 K for bare carbon fibers (CFs) and the ACFs to be 97 and 260-285 mJ·m-2, respectively, which lie in the regime of secondary bonding of physical adsorption. Our analysis indicates that these are impacted by micropores and defects on the carbon surfaces. Comparing the γSL obtained by the traditional Gray's method, our method is concluded as the most accurate and reliable value for the hydrophobic dispersive surface component of porous carbonaceous materials. As such, it could serve as a valuable tool in designing interface engineering in adsorption-related applications.

Surface energetics of graphene oxide and reduced graphene oxide determined by inverse gas chromatographic technique at infinite dilution at room temperature.

Graphene is of great interest for many far-reaching applications that involve interparticle interactions in adsorbents, coatings, and composites. A deep understanding of the surface components has been crucial but achieving the most accurate and reliable values of these, unaffected by experimental conditions or the analytical techniques used, remains a major challenge. To this end, we have proposed in this paper a novel approach for the first time, to the best of our knowledge, to determine London dispersive and specific (polar) components including the Lewis acid-base character of the surface free energy of graphene materials (graphene oxide (GO), reduced graphene oxide (rGO), and graphite) using inverse gas chromatography (IGC) technique at an infinite dilution. We have estimated the London dispersive surface energy values of graphite, GO, and rGO as van der Waals interaction to be 156-179, 89-106, and 110-119 mJ m-2, respectively, in the temperature range of 320-360 K. These are attributable to the surface properties impacted by the oxygen moieties, defects, and micropores on the carbon frameworks. Further, the acceptor (KA) and donor (KD) parameters of GO were found to be 0.71 and 0.96, respectively, while those of rGO were 0.54 and 1.05. Notably, the GO is more of the Lewis acid character that could be amphoteric, while the Lewis base characteristics of both GO and rGO are not significantly changed. These results provide foundational knowledge to understand the physicochemical properties of graphene surfaces, which should be helpful to designing interface engineering in various applications.

Optimization of the droplet electroporation method for wild type Chlamydomonas reinhardtii transformation.

We performed the transformation of a wild type Chlamydomonas reinhardtii by optimizing previously developed droplet EP method. For more effective and faster optimization, we used DNA dying fluorescent molecule (Yo-Pro-1) for finding optimal EP conditions instead of using protein expression based evaluation method. By examining wider range of electrical parameter space together with the analysis of total current flow of EP process, we found optimal EP conditions. The obtained optimal EP conditions were verified by CFP transgene expression experiments. By applying the optimal EP conditions to the transformation of C. reinhardtii, we obtained transformants and analyzed them using PCR. Finally, implications and future work are discussed.

A continuous droplet electroporation system for high throughput processing.

A continuous droplet electroporation (EP) system capable of handling a billion cells has been proposed and demonstrated using a proof-of-concept prototype design. Numerical simulations were conducted to design the new system and to compare the continuous droplet EP system with the previous single droplet EP system. Through parametric studies on the applied voltage and flow rate, a much higher cyan fluorescent protein transgene expression efficiency (38.8 ± 8.9%) was accomplished over that of the previous single droplet EP system. A parallel continuous droplet EP system is also demonstrated by introducing additional electrode pairs into the continuous droplet EP system to achieve ultrahigh throughput. Finally, the significance and meaning of the present work and future development direction have been discussed.

Abolishment of N-glycan mannosylphosphorylation in glyco-engineered Saccharomyces cerevisiae by double disruption of MNN4 and MNN14 genes.

Mannosylphosphorylated glycans are found only in fungi, including yeast, and the elimination of mannosylphosphates from glycans is a prerequisite for yeast glyco-engineering to produce human-compatible glycoproteins. In Saccharomyces cerevisiae, MNN4 and MNN6 genes are known to play roles in mannosylphosphorylation, but disruption of these genes does not completely remove the mannosylphosphates in N-glycans. This study was performed to find unknown key gene(s) involved in N-glycan mannosylphosphorylation in S. cerevisiae. For this purpose, each of one MNN4 and five MNN6 homologous genes were deleted from the och1Δmnn1Δmnn4Δmnn6Δ strain, which lacks yeast-specific hyper-mannosylation and the immunogenic α(1,3)-mannose structure. N-glycan profile analysis of cell wall mannoproteins and a secretory recombinant protein produced in mutants showed that the MNN14 gene, an MNN4 paralog with unknown function, is essential for N-glycan mannosylphosphorylation. Double disruption of MNN4 and MNN14 genes was enough to eliminate N-glycan mannosylphosphorylation. Our results suggest that the S. cerevisiae och1Δmnn1Δmnn4Δmnn14Δ strain, in which all yeast-specific N-glycan structures including mannosylphosphorylation are abolished, may have promise as a useful platform for glyco-engineering to produce therapeutic glycoproteins with human-compatible N-glycans.

Increased mannosylphosphorylation of N-glycans by heterologous expression of YlMPO1 in glyco-engineered Saccharomyces cerevisiae for mannose-6-phosphate modification.

Mannosylphosphorylated N-glycans found in yeasts can be converted to those containing mannose-6-phosphate, which is a key factor for lysosomal targeting. In the traditional yeast Saccharomyces cerevisiae, both ScMNN4 and ScMNN6 genes are required for efficient mannosylphosphorylation. ScMnn4 protein has been known to be a positive regulator of ScMnn6p, a real enzyme for mannosylphosphorylation. On the other hand, YlMpo1p, a ScMnn4p homologue, mediates mannosylphosphorylation in Yarrowia lypolytica without the involvement of ScMnn6p homologues. In this study, we show that heterologous expression of YlMpo1p can perform and enhance mannosylphosphorylation in S. cerevisiae in the absence of ScMnn4p and ScMnn6p. Moreover, the level of mannosylphosphorylation of N-glycans enhanced by YlMpo1p overexpression is much higher than that with ScMnn4p overexpression, and this is highlighted further in Scmnn4- and Scmnn6-disrupted mutants. When YlMpo1p overexpression is applied to glyco-engineered S. cerevisiae in which the synthesis of immunogenic glycans is abolished, a great increase of bi-mannosylphosphorylated glycan is observed. Through an in vitro process involving the uncapping of the outer mannose residue, this bi-mannosylphosphorylated structure is changed to a bi-phosphorylated structure with high affinity for mannose-6-phosphate receptor. The superior ability of YlMpo1p to increase bi-mannosylphosphorylated glycan in yeast shows promise for the production of therapeutic enzymes with improved lysosomal targeting capability.

Effect of polarizable lone pair cations on the second-harmonic generation (SHG) properties of noncentrosymmetric (NCS) Bi(2-x)Y(x)TeO5 (x = 0-0.2).

Y(3+)-doped noncentrosymmetric (NCS) bismuth tellurite materials, Bi(2-x)Y(x)TeO5 (x = 0, 0.1, and 0.2), have been synthesized through standard solid-state reactions and structurally characterized by powder neutron diffraction. The reported NCS materials crystallize in the orthorhombic space group Abm2 (no. 39), and exhibit pseudo-three-dimensional frameworks that are composed of BiO3, BiO5, and TeO3 polyhedra. Detailed diffraction studies show that the cell volume of Bi(2-x)Y(x)TeO5 decreases with an increasing amount of Y(3+)on the Bi(3+) sites. However, no ordering between Bi(3+) and Y(3+) was observed in the Bi(2-x)Y(x)TeO5. Powder second-harmonic generation (SHG) measurements, using 1064 nm radiation, reveal that Bi2TeO5, Bi(1.9)Y(0.1)TeO5, and Bi(1.8)Y(0.2)TeO5 exhibit SHG efficiencies of approximately 300, 200, and 60 times that of α-SiO2, respectively. The reduction in SHG for Y(3+)-doped materials is consistent with the lack of net moment originating from polyhedra with a polarizable Bi(3+) cation.

Strong second harmonic generation (SHG) originating from combined second-order Jahn-Teller (SOJT) distortive cations in a new noncentrosymmetric tellurite, InNb(TeO4)2.

Pure polycrystalline bulk samples and single crystals of a novel polar noncentrosymmetric (NCS) indium niobium tellurium oxide, InNb(TeO4)2, have been synthesized through solid-state reactions with In2O3, Nb2O5, and TeO2. InNb(TeO4)2 exhibits a three-dimensional structural backbone consisting of layers of corner-shared InO6 octahedra, layers of corner-shared NbO6 octahedra, and chains of corner-shared TeO4 polyhedra. The infrared spectrum shows In-O, Nb-O, and Te-O vibrations, and the UV-vis diffuse reflectance spectrum shows a band gap of 3.5 eV for the reported material. Thermal analysis indicates that InNb(TeO4)2 is thermally stable to 740 °C. Powder second-order nonlinear optical (NLO) measurements reveal that InNb(TeO4)2 has a strong second harmonic generation (SHG) efficiency, 100 times that of α-SiO2. A detailed structural examination indicates that the strong SHG is due to the combined net polarization originating from the distorted environment of second-order Jahn-Teller (SOJT) cations such as Nb(5+) and Te(4+). Elemental analysis, the magnitude of distortion, and calculations of dipole moments for InNb(TeO4)2 are also presented.

Noncentrosymmetric YVSe2O8 and centrosymmetric YVTe2O8: macroscopic centricities influenced by the size of lone pair cation linkers.

Two new quaternary vanadium selenite and tellurite, i.e., YVSe2O8 and YVTe2O8, have been synthesized through hydrothermal and solid-state reactions. Both of the reported materials exhibit three-dimensional framework structures that are composed of layers of corner-shared VO6 octahedra, layers of edge-shared YO8 groups, and SeO3 or TeO3 linkers. While YVSe2O8 crystallizes in the orthorhombic noncentrosymmetric (NCS) space group, Abm2, YVTe2O8 reveals the monoclinic centrosymmetric (CS) space group, C2/m. The band gaps for YVSe2O8 and YVTe2O8 are calculated to be 2.7 and 2.2 eV, respectively, from the UV-vis diffuse reflectance spectra. Powder second harmonic generation (SHG) measurements using 1064 nm radiation reveals that NCS YVSe2O8 has a similar SHG efficiency to that of NH4H2PO4 (ADP). Detailed explanations of the structure-property relationships, effect of lone pair cation size on the macroscopic centricities, and full characterizations including infrared spectroscopy, thermal analyses, and elemental analyses are also presented.

α-ScVSe2O8, ß-ScVSe2O8, and ScVTe2O8: new quaternary mixed metal oxides composed of only second-order Jahn-Teller distortive cations.

Three new quaternary scandium vanadium selenium/tellurium oxides, α-ScVSe2O8, ß-ScVSe2O8, and ScVTe2O8 have been synthesized through hydrothermal and standard solid-state reactions. Although all three reported materials are stoichiometrically similar, they exhibit different crystal structures: α-ScVSe2O8 has a three-dimensional framework structure consisting of ScO6, VO6, and SeO3 groups. ß-ScVSe2O8 reveals another three-dimensional framework composed of ScO7, VO5, and SeO3 polyhedra. ScVTe2O8 shows a layered structure with ScO6, VO4, and TeO4 polyhedra. Interestingly, the constituent cations, that is, Sc(3+), V(5+), Se(4+), and Te(4+) are all in a distorted coordination environment attributable to second-order Jahn-Teller (SOJT) effects. Complete characterizations including infrared spectroscopy, elemental analyses, thermal analyses, dipole moment calculation, and the magnitudes of out-of-center distortions for the compounds are reported. Transformation reactions suggest that α-ScVSe2O8 may change to ß-ScVSe2O8, and then to Sc2(SeO3)3·H2O under hydrothermal conditions.

Promotion of both proliferation and neuronal differentiation in pluripotent P19 cells with stable overexpression of the glutamine transporter slc38a1.

BACKGROUND: We previously demonstrated the functional expression in newborn rat neocortical astrocytes of glutamine transporter (GlnT = slc38a1) believed to predominate in neurons over astroglia in the brain. In order to evaluate the possible role of this transporter in neurogenesis, we attempted to establish stable transfectants of GlnT in mouse embryonal carcinoma P19 cells endowed to proliferate for self-renewal and differentiate into progeny cells such as neurons and astroglia, in addition to in vitro pharmacological profiling of the green tea ingredient theanine, which is shown to be a potent inhibitor of glutamine transport mediated by GlnT in cultured neurons and astroglia. METHODOLOGY/PRINCIPAL FINDINGS: The full-length coding region of rat GlnT was inserted into a vector for gene transfection along with selection by G418, followed by culture with all-trans retinoic acid under floating conditions and subsequent dispersion for spontaneous differentiation under adherent conditions. Stable overexpression of GlnT led to marked increases in the size of round spheres formed during the culture for 4 days and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide reduction, with concomitant promotion of subsequent differentiation into cells immunoreactive for a neuronal marker protein. In these stable GlnT transfectants before differentiation, drastic upregulation was seen for mRNA expression of several proneural genes with a basic helix-loop-helix domain such as NeuroD1. Although a drastic increase was seen in NeuroD1 promoter activity in stable GlnT transfectants, theanine doubled NeuroD1 promoter activity in stable transfectants of empty vector (EV), without affecting the promoter activity already elevated in GlnT transfectants. Similarly, theanine promoted cellular proliferation and neuronal differentiation in stable EV transfectants, but failed to further stimulate the acceleration of both proliferation and neuronal differentiation found in stable GlnT transfectants. CONCLUSIONS/SIGNIFICANCE: GlnT would promote both proliferation and neuronal differentiation through a mechanism relevant to the upregulation of particular proneural genes in undifferentiated P19 cells.

Promoted neuronal differentiation after activation of alpha4/beta2 nicotinic acetylcholine receptors in undifferentiated neural progenitors.

BACKGROUND: Neural progenitor is a generic term used for undifferentiated cell populations of neural stem, neuronal progenitor and glial progenitor cells with abilities for proliferation and differentiation. We have shown functional expression of ionotropic N-methyl-D-aspartate (NMDA) and gamma-aminobutyrate type-A receptors endowed to positively and negatively regulate subsequent neuronal differentiation in undifferentiated neural progenitors, respectively. In this study, we attempted to evaluate the possible functional expression of nicotinic acetylcholine receptor (nAChR) by undifferentiated neural progenitors prepared from neocortex of embryonic rodent brains. METHODOLOGY/PRINCIPAL FINDINGS: Reverse transcription polymerase chain reaction analysis revealed mRNA expression of particular nAChR subunits in undifferentiated rat and mouse progenitors prepared before and after the culture with epidermal growth factor under floating conditions. Sustained exposure to nicotine significantly inhibited the formation of neurospheres composed of clustered proliferating cells and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide reduction activity at a concentration range of 1 µM to 1 mM without affecting cell survival. In these rodent progenitors previously exposed to nicotine, marked promotion was invariably seen for subsequent differentiation into cells immunoreactive for a neuronal marker protein following the culture of dispersed cells under adherent conditions. Both effects of nicotine were significantly prevented by the heteromeric α4ß2 nAChR subtype antagonists dihydro-ß-erythroidine and 4-(5-ethoxy-3-pyridinyl)-N-methyl-(3E)-3-buten-1-amine, but not by the homomeric α7 nAChR subtype antagonist methyllycaconitine, in murine progenitors. Sustained exposure to nicotine preferentially increased the expression of Math1 among different basic helix-loop-helix proneural genes examined. In undifferentiated progenitors from embryonic mice defective of NMDA receptor subunit-1, nicotine was still effective in significantly inhibiting the proliferation. CONCLUSIONS/SIGNIFICANCE: Functional α4ß2 nAChR subtype would be constitutively expressed to play a role in the mechanism underlying the determination of proliferation and subsequent differentiation fate into a neuronal lineage in association with preferential promotion of Math1 expression in undifferentiated neural progenitors of developing rodent neocortex independently of NMDA receptor activation.

Expression of thioredoxin during progression of hamster and human cholangiocarcinoma.

Thioredoxin (Trx) is a multifunctional redox protein that has growth-promoting and anti-apoptotic effects on cells and protects cells from endogenous and exogenous free radicals. Recently, altered expression of Trx has been reported in various cancers. In the present study, we investigated altered expression of Trx at the precancerous and carcinogenic phases during cholangiocarcinogenesis in a hamster cholangiocarcinoma (ChC) model, using semiquantitative immunohistochemical and Western blot analyses. Moreover, to determine if the results correlated well with those in human ChCs, we carried out a comparative immunohistochemical study for Trx in tissue-arrayed human ChCs with different grades of tumor cell differentiation. Trx was found highly expressed in the cytoplasm of dysplastic bile ducts with highly abnormal growth patterns and ChCs irrespective of tumor type or tumor cell differentiation. Overexpression of Trx at the precancerous and carcinogenic phases was further supported by significant elevation of Trx protein in Western blotting. The results from the hamster ChCs were in good agreement with those from human ChCs. Our results strongly suggested that the redox regulatory function of Trx plays an important role in bile duct cell transformation and tumor progression during cholangiocarcinogenesis.

Primary subcutaneous leiomyosarcoma of the hamster hind leg.

We report the first case of a primary subcutaneous leiomyosarcoma that originated in the hind leg of a hamster and metastasized to the bone marrow, lung and diaphragm. A 10-month-old female Syrian golden hamster was presented with a large, firm, white subcutaneous mass in the right hind leg. The tumor invaded into the bone marrow and small nodules were also present in the lung and diaphragm; however, no tumor masses were found in the visceral organs. Histologically, the tumors were spindle cell sarcomas, composed of densely packed pleomorphic spindle cells with oval to elongate nuclei and moderate amounts of eosinophilic cytoplasm. Immunohistochemically, the neoplastic cells were positive for vimentin, desmin, and smooth muscle actin, but negative for alpha-sarcomeric actin and S-100. Thus, the diagnosis was primary leiomyosarcoma of the hind leg with metastasis to the bone marrow, lung and diaphragm. To the best of our knowledge, this is the first report of spontaneous primary subcutaneous leiomyosarcoma of the hind leg with distant metastasis in laboratory animals.